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      E6-Specific Detection and Typing of Human Papillomaviruses in Oral Cavity Specimens from Iranian Patients

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          Abstract

          Background:

          Detection and quantification of human Papillomavirus (HPV) genome in oral carcinoma play an important role in diagnosis, as well as implications for progression of disease.

          Methods:

          We evaluated tissues from 50 esopharyngeal cancers collected from different regions of Iran for HPV E6 using the two type-specific primers sets. E6 gene of HPV genotypes was amplified by specific primers. The sensitivity of PCR assay was analyzed and determined using HPV-DNA-containing plasmids. Real-time PCR was utilized to determine the prevalence and HPV viral load in patients with oral cavity squamous cell carcinoma.

          Results:

          Eighteen (36%) specimens were positive for HPV. Among the 18 positive specimens, 10 showed HPV-18 (55.55%), and 8 specimens were positive for HPV-11 (44.44%). Of the 18 infected specimens, 6 (33.32%) and 12 (66.65%) were identified as high-titer and low-titer viral load, respectively.

          Conclusions:

          The PCR-based assay, developed in the current study, could be used for HPV detection, quantification, and genotyping in epidemiological and clinical studies.

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          Oral squamous cell carcinoma: review of prognostic and predictive factors.

          Frederico S. Regateiro, Luisa Ferreira, Gustavo Januário … (2006)
          Oral squamous cell carcinoma has a remarkable incidence worldwide and a fairly onerous prognosis, encouraging further research on factors that might modify disease outcome. In this review article, the authors approach the factors that may exert influence on the prognosis and eventually guide the selection of patients for more aggressive therapies. Published scientific data was collected, selected, and grouped into 3 main clusters: patient-, tumor-, and treatment-related factors. Well established aspects are discussed, but also those less common or with only supposed usefulness. Disease staging, extracapsular dissemination, resection margin free of disease, and tumor thickness are factors with high influence on the prognosis. There has been an increasing interest in the study of tumor molecular factors, and some have been strongly correlated with the outcome, showing promising pathways for the future development of more effective prognosis systems and anticancer therapies.
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            GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high-throughput identification of human papillomavirus genotypes.

            Adriaan van den Brule, René Pol, Nathalie Fransen-Daalmeijer … (2002)
            In this study, we developed a simple and fast typing procedure for 37 mucosotropic human papillomavirus (HPV) types using a nonradioactive reverse line blotting (RLB) procedure for general primer (GP5+/6+) PCR products. This system has the advantages not only that in a simple format, up to 42 PCR products can be simultaneously typed per membrane per day, but also that after stripping, the membranes can be easily rehybridized at least 15 times without a loss of signal. RLB appeared highly specific, and its sensitivity was identical to that of conventional typing performed with type-specific oligonucleotide probes in an enzyme immunoassay (EIA). The performance of RLB typing was evaluated with samples of HPV-positive cervical scrapings (n = 196) and biopsies of cervical premalignant lesions (n = 100). The distribution of HPV genotypes detected in these samples was in line with the distribution expected on the basis of literature data. In addition, RLB and EIA typing procedures were compared for the typing of high-risk HPV types in GP5+/6+ PCR products of 210 cervical scrapings from high-risk HPV-positive women who participated in a population-based screening program. The typing procedures had an excellent overall agreement rate of 96.5% (kappa value, 0.77). RLB was successful in detecting multiple HPV infections as well as single infections. In conclusion, the GP5+/6+ PCR-RLB procedure appeared to be a reliable and simple approach that may be of great value for large epidemiological studies, population-based cervical cancer screening programs, and vaccination trials that require high-throughput HPV typing.
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              Detection and typing of human papillomavirus by e6 nested multiplex PCR.

              K Sotlar, D Diemer, A Dethleffs … (2004)
              A nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and type-specific primers was evaluated for the detection and typing of human papillomavirus (HPV) genotypes 6/11, 16, 18, 31, 33, 35, 39, 42, 43, 44, 45, 51, 52, 56, 58, 59, 66, and 68 using HPV DNA-containing plasmids and cervical scrapes (n = 1,525). The performance of the NMPCR assay relative to that of conventional PCR with MY09-MY11 and GP5+-GP6+ primers, and nested PCR with these two primer sets (MY/GP) was evaluated in 495 cervical scrapes with corresponding histologic and cytologic findings. HPV prevalence rates determined with the NMPCR assay were 34.7% (102 of 294) in the absence of cervical intraepithelial neoplasia (CIN 0), 94.2% (113 of 120) in the presence of mild or moderate dysplasia (CIN I/II), and 97.8% (44 of 45) in the presence of severe dysplasia (CIN III). The combination of all four HPV detection methods applied in the study was taken as "gold standard": in all three morphological subgroups the NMPCR assay had significantly (P < 0.0001) higher sensitivities than the MY09-MY11 and GP5+-GP6+ assays and sensitivities comparable or equal to those of the MY/GP assay. All 18 HPV genotypes investigated were detected among the clinical samples. The ratio of high- to low-risk HPV genotypes increased from 4:1 (80 of 103) in CIN 0 to 19:1 (149 of 157) in CIN I to III. Multiple infections were detected in 47.9% (124 of 259) of the patients. In conclusion, the novel NMPCR method is a sensitive and useful tool for HPV DNA detection, especially when exact HPV genotyping and the identification of multiple HPV infections are required.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Iran Biomed J
                Journal ID (iso-abbrev): Iran. Biomed. J
                Title: Iranian Biomedical Journal
                Publisher: Pasteur Institute (Iran )
                ISSN (Print): 1028-852X
                ISSN (Electronic): 2008-823X
                Publication date (Print): November 2017
                Volume: 21
                Issue: 6
                Pages: 411-416
                Affiliations
                [1 ]Laboratory Science Research Center, Golestan University of Medical Sciences, Gorgan, Iran
                [2 ]Department of Microbiology, Faculty of Medicine, Golestan University of Medical Sciences, Gorgan, Iran
                [3 ]Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
                [4 ]Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
                [5 ]Hepatitis Research Center, Khorramabad University of Medical Sciences, Lorestan, Iran
                [6 ]Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran
                Author notes
                Corresponding Author: Mehrdad Ravanshad Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. P.O. Box. 14115-111. Tel.: (+98-21) 82883836; Fax: (+98-21) 82883836; E-mail: hadirazavi96@ 123456gmail.com
                Article
                Publisher ID: IBJ-21-411
                DOI: 10.18869/acadpub.ibj.21.6.411
                PMC ID: 5572438
                PubMed ID: 28460428
                SO-VID: 76d2f26b-c571-4f5c-b7b9-fa492e4380d3
                Copyright © Copyright: © Iranian Biomedical Journal
                License:

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, ( http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Date received : 24 December 2016
                Date revision received : 31 December 2016
                Date accepted : 26 January 2017
                Categories
                Subject: Short Communication

                Keywords: real-time pcr,genotyping,iran
                Data availability:
                Keywords: real-time pcr, genotyping, iran

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